The major objective of the proposed investigation is to define and characterize several of the enzymes that are involved in the synthesis of adenovirus 2 (Ad 2) DNA during productive infection in human KB cells. The study will also attempt to dilineate the specific function of the DNA and RNA polymerases responsible for the replication of the Ad 2 genome. The research will entail the use of a soluble, multienzyme DNA synthesizing complex which is isolated from the nuclei of Ad 2 infected KB cells late after infection. The demonstration of various enzyme activities can be accomplished in both the intact complex and a soluble preparation after removal of the endogenous DNA template. The complex synthesizes exclusively Ad 2 DNA in vitro and this small, well-characterized genome lends itself readily to standard biochemical analyses. Since adenovirus uses cellular DNA and RNA polymerases for its own replication, this system is proposed as a model for the study of the enzymology and specific processes involved in the replication of mammalian cell genomes much as the study of phage replication has served as a model for the replication of bacterial chromosomes. Since adenovirus causes tumors in lower animals and human KB cells were from a nasopharyngeal carcinoma, this study relates to cancer problems. The specific aims of the proposed research are: (a) to purify and characterize the DNA polymerase(s) in the soluble multienzyme complex, (b) to determine the specific function of the DNA polymerase(s) in the complex, (c) to purify and characterize the RNA polymerase activity in the complex, and (d) to demonstrate and analyze the interaction of the DNA polymerase(s) in the replication complex with the RNA product of the RNA polymerase also present in the complex.